基于NF-κB信号通路探讨湿热痹颗粒治疗强直性脊柱炎湿热证小鼠的作用机制OA

Objective:To evaluate the therapeutic efficacy of Shirebi granules(SG)in a murine model of ankylosing spon-dylitis with damp-heat syndrome(ASDH)and to elucidate the underlying molecular mechanisms.Methods:An ASDH mouse model was established by inducing ankylosing spon-dylitis with proteoglycan and administering oil and rice wine orally,while exposing the mice to an artificial climate cham-ber to simulate damp-heat syndrome.Mice were randomly assigned to five groups:blank control group,model group,low and high doses of SG(2.5 and 5.0 g/kg,respectively)group,and a positive control group treated with sulfasalazine(50 mg/kg).Treatments were administered orally for six weeks.Quantitative assessment of traditional Chinese medicine(TCM)syndromes associated with damp-heat was carried out,and arthritis evaluation was performed on the limb joints for each group of mice before administration and at 2,4,and 6 weeks after administration.HE staining was employed to assess the histopathology of spinal joint sections.Serum levels of TNF-α,IL-1β,IL-4,and IL-10 were quantified via ELISA.Gene expression of IκBα and NF-κB p65 in spinal joint tissues was assessed using RT-qPCR.Western Blot analysis was performed to determine the relative protein expression of p-IκBα/IκBα and p-p65/p65.Results:Administration of both high and low doses of SG ameliorated clinical manifestations associat-ed with damp-heat syndrome,including reduced activity,lethargy,irritability,dull fur,loose stools,and yellowish malodor-ous urine.Quantitative TCM syndrome scores were significantly lower in the treatment group than in the model group at weeks 4 and 6 post-treatment(P＜0.05).Arthritis scores in the treatment groups exhibited a more gradual progression over time relative to the model group and were significantly lower at both 4 and 6 weeks(P＜0.05).Both dosages of SG groups demonstrated a significant reduction in pathological lesions within the spinal joints,including decreased inflamma-tory cell infiltration,mesenchymal cell proliferation,and excessive matrix formation.Correspondingly,histopathological scores in these groups were significantly lower than those in the model group(P＜0.05).Both dosages of SG significantly decreased serum concentrations of TNF-α and IL-1β compared to the model group(P＜0.05).Furthermore,compared to both the model group and the positive control group,treatment with SG at these dosages led to a significant elevation in the levels of IL-4 and IL-10(P＜0.05).At the molecular level,treatment significantly downregulated mRNA expression of IκBα and NF-κB p65 in spinal joint tissues(P＜0.05)and reduced the protein expression ratios of p-IκBα/IκBα and p-p65/p65(P＜0.05).Conclusion:SG effectively mitigate TCM syndrome manifestations and joint pathology in ASDH mice.The therapeutic effects are associated with modulation of the balance between pro-inflammatory and anti-inflamma-tory cytokines and suppression of NF-κB signaling pathway activation.